Immunofluorescence Using the ibidi Chambers
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Bottom material |
Standard glass slide |
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Additional coverslips required? |
No |
No |
Yes (for mounting) |
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Microscope type |
Inverted |
Inverted |
Inverted & upright |
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Mounting medium |
Non-hardening |
Non-hardening |
Hardening |
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Sample storage |
Short-term |
Short-term |
Long-term |
Benefits of ibidi µ-Chambers
- High-resolution imaging
The ibidi slides are ideal for widefield fluorescence, confocal imaging, FRAP, FRET, FLIM, and undisturbed phase contrast imaging. - Fast and simple handling
The all-in-one chambers simplify your immunofluorescence protocol. - Cost-effective experiments
Only a small number of cells and low reagent volumes are needed.
Comparison of Immunocytochemistry Protocols:
Traditional Staining vs. Staining With ibidi Solutions
When using any of the ibidi solutions, the immunofluorescence staining protocol is much shorter than the traditional protocol. There is no need to grow the cells on loose coverslips—the cells can be grown and stained directly in the ibidi slides.
Protocol With Cells on Coverslips
(Traditional method with nail polish mounting)
- Sterilize coverslips and slides
- Coat coverslips
- Place sterile coverslips into 6-well plate
- Seed cells in large volume
- Peel off the coverslip
- Wash
- Fix – wash – permeabilize – wash – block
- Incubate in primary antibody – wash – incubate in secondary antibody
- Wash
- Mount cells with mounting medium
- Mount coverslip with nail polish
Protocol With ibidi µ-Slides
(Time-saving method using all-in-one chambers)
- Sterilize coverslips and slides
- Coat coverslips
- Place sterile coverslips into 6-well plate
- Seed cells in large volume
- Peel off the coverslip
- Wash
- Fix – wash – permeabilize – wash – block
- Incubate in primary antibody – wash – incubate in secondary antibody
- Wash
- Mount cells with mounting medium
- Mount coverslip with nail polish
The ibidi Mounting Medium and the ibidi Mounting Medium With DAPI have a very low autofluorescence, prevent photobleaching and allow the sample to be stored for several weeks on the µ-Slide without the need for additional coverslips.
If your experiment requires the long-term storage of immunostained samples, we recommend using the ibidi Chamber Slides, removable.
Find detailed immunofluorescence staining protocols in the ibidi Application Notes:
- AN 02: Fluorescence Staining using a µ-Slide I (PDF)
- AN 09: Fluorescence Staining using a µ-Slide VI 0.4 (PDF)
- AN 15: Fluorescence Staining using a µ-Slide y-shaped (PDF)
- AN 16: Fluorescence Staining Using the µ-Slide 8 Well high (PDF)
- AN 49: Fluorescence Staining using a 12 Well Chamber, removable (PDF)
- AN 50: Fluorescence Staining using a 3 Well Chamber, removable (PDF)
- AN 45: Mounting Medium Types (PDF)
Read on and see some Experimental Examples for immunofluorescence, or learn more about a typical Workflow of an Immunofluorescence Staining.